Korean J Hepatobiliary Pancreat Surg.  2015 May;19(2):47-58. 10.14701/kjhbps.2015.19.2.47.

Toward angiogenesis of implanted bio-artificial liver using scaffolds with type I collagen and adipose tissue-derived stem cells

• Lee JG
• Bak SY
• Nahm JH
• Lee SW
• Min SO
• Kim KS

• KMID: 2243049
• DOI: http://doi.org/10.14701/kjhbps.2015.19.2.47

Abstract

BACKGROUNDS/AIMS
Stem cell therapies for liver disease are being studied by many researchers worldwide, but scientific evidence to demonstrate the endocrinologic effects of implanted cells is insufficient, and it is unknown whether implanted cells can function as liver cells. Achieving angiogenesis, arguably the most important characteristic of the liver, is known to be quite difficult, and no practical attempts have been made to achieve this outcome. We carried out this study to observe the possibility of angiogenesis of implanted bio-artificial liver using scaffolds.
METHODS
This study used adipose tissue-derived stem cells that were collected from adult patients with liver diseases with conditions similar to the liver parenchyma. Specifically, microfilaments were used to create an artificial membrane and maintain the structure of an artificial organ. After scratching the stomach surface of severe combined immunocompromised (SCID) mice (n=4), artificial scaffolds with adipose tissue-derived stem cells and type I collagen were implanted. Expression levels of angiogenesis markers including vascular endothelial growth factor (VEGF), CD34, and CD105 were immunohistochemically assessed after 30 days.
RESULTS
Grossly, the artificial scaffolds showed adhesion to the stomach and surrounding organs; however, there was no evidence of angiogenesis within the scaffolds; and VEGF, CD34, and CD105 expressions were not detected after 30 days.
CONCLUSIONS
Although implantation of cells into artificial scaffolds did not facilitate angiogenesis, the artificial scaffolds made with type I collagen helped maintain implanted cells, and surrounding tissue reactions were rare. Our findings indicate that type I collagen artificial scaffolds can be considered as a possible implantable biomaterial.

Keyword

Tissue scaffolds; Liver; Artificial; Neovascularization; Physiologic; Biocompatible materials

MeSH Terms

Actin Cytoskeleton
Adult
Animals
Artificial Organs
Biocompatible Materials
Collagen Type I*
Humans
Liver Diseases
Liver*
Membranes, Artificial
Mice
Stem Cells*
Stomach
Tissue Scaffolds
Vascular Endothelial Growth Factor A
Biocompatible Materials
Collagen Type I
Membranes, Artificial
Vascular Endothelial Growth Factor A

Figure

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  Fig. 1 Experimental model.

• 

  Fig. 2 Schematic presentation and a photograph of the artificial scaffold model.

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  Fig. 3 Adhesion of artificial scaffold to surrounding organs 30 days after implantation into SCID mice. (A) View of internal organs after opening the mouse abdomen. (B) The artificial scaffold (black arrow) was exposed with surgical forceps. The omentum was covering the artificial scaffold.

• 

  Fig. 4 H&E-stained images of the artificial scaffold material processed 30 days after implantation into SCID mice. Red eosin stain showed the artificial collagen scaffold. Black arrows indicate where the scaffold was sutured to the mouse stomach.

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  Fig. 5 H&E-stained collagen of the artificial scaffold 30 days after implantation into SCID mice. The area in the black circle (A) is magnified in (B-D). (D) The ×200 magnification shows the lack of nuclei in the red eosin-stained collagen surrounding the artificial scaffold.

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  Fig. 6 Magnification of the inner side of the H&E-stained artificial scaffold 30 days after implantation into SCID mice. The area in the black circle (A) is magnified in (B-D). (D) The ×200 magnification shows eosin-stained collagen and hematoxylin-stained adipose tissue-derived stem cells within the artificial scaffold.

• 

  Fig. 7 A VEGF-stained artificial scaffold processed 30 days after implantation into SCID mice. The area in the black circle (A) is magnified. (D) ×100 and (F) ×200 magnifications of the area in the black dotted circle shown in (B). The cells inside the artificial scaffold include hematoxylin-stained adipose tissue-derived stem cells and diaminobenzidine-stained collagen. (C) ×100 magnification and (E) ×200 magnification of the area in the black circle shown in (B). The outside of the artificial scaffold contained mouse omentum cells with no collagen. There was no endothelial or vascular structure or VEGF expression.

• 

  Fig. 8 A CD34-stained artificial scaffold processed 30 days after implantation into SCID. The area in the black circle (A) is magnified. (D) ×100 and (F) ×200 magnifications of the area in the black dotted circle area (B). The cells inside the artificial scaffold include hematoxylin-stained adipose tissue-derived stem cells and diaminobenzidine-stained collagen. (C) ×100 magnification and (E) ×200 magnification of the area in the black circle shown in (B). The outside of the artificial scaffold contained mouse omentum cells with no collagen. There was no endothelial or vascular structure or CD34 expression.

• 

  Fig. 9 A CD105-stained artificial scaffold processed 30 days after implantation into SCID. The area in the black circle (A) is magnified. (D) ×100 and (F) ×200 magnifications of the area in the black dotted circle area (B). The cells inside the artificial scaffold include hematoxylin-stained adipose tissue-derived stem cells and diaminobenzidine-stained collagen. (C) ×100 magnification and (E) ×200 magnification of the area in the black circle shown in (B). The outside of the artificial scaffold contained mouse omentum cells with no collagen. There was no endothelial or vascular structure or CD105 expression.

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